Sequencing of cDNA clones that encode bovine ferritin H and L chains.
The molecular weight of the liver-type subunit (L) of bovine ferritin is much larger than that of the heart-type subunit (H) as determined by SDS-PAGE (L, 20.5 kDa; H, 18.4 kDa). The migration of these two subunits on SDS-PAGE gels, relative to each other, is opposite to that reported for ferritin L and H subunits in other mammalian species (L, 19 kDa; H, 21 kDa). To determine the cause of this anomaly, full-length cDNA clones of the bovine L and H chains were isolated from a bovine spleen gamma gt11 cDNA library and sequenced. The amino acid sequences of the L and H chains of bovine ferritin, deduced from their cDNA sequences, contained open reading frames coding for 174 and 180 amino acid residues with calculated molecular weights of 19,856 and 20,920 Da, respectively. The deduced amino acid sequence of the L chain shows 86%, 84%, 87%, 83% and 83% homology with the amino acid sequences of horse, human, rabbit, rat and mouse L chains, respectively. The H chain displays a higher homology with the human, rat and mouse H chains (91%, 92% and 93%, respectively). In addition, the bovine L chain did not contain the extra octapeptide present in rodent L chains, and bovine, L and H chains did not react with concanavalin A. The bovine L and H chains expressed using a baculovirus expression system showed almost the same mobilities as those of bovine spleen ferritin, respectively, by SDS-PAGE. These results suggest that the much slower mobility of the bovine L chain compared with other mammalian L chains on SDS-PAGE cannot be attributed to insertion(s) of amino acid(s) or peptide(s) into the L chain, to the deletion(s) of them of it or to the addition of carbohydrate chains(s) but may result from significant differences in the binding affinity of SDS for bovine ferritin L chains.
Orino
K
K
Laboratory of Biochemistry, School of Veterinary Medicine and Animal Sciences, Kitasato University, Aomori, Japan. orino@vmas.kitasato-u.ac.jp
Eguchi
K
K
Nakayama
T
T
Yamamoto
S
S
Watanabe
K
K
D000595:Amino Acid Sequence; D000818:Animals; D001052:Apoferritins; D001483:Base Sequence; D002417:Cattle; D003001:Cloning, Molecular; D017124:Conserved Sequence; D018076:DNA, Complementary / Q000737:chemistry; D004591:Electrophoresis, Polyacrylamide Gel; D005293:Ferritins / Q000235:genetics* / Q000276:immunology; D006801:Humans; D051379:Mice; D008969:Molecular Sequence Data; D009206:Myocardium / Q000737:chemistry; D011817:Rabbits; D051381:Rats; D017421:Sequence Analysis; D017386:Sequence Homology, Amino Acid; D013154:Spleen / Q000737:chemistry
D018076:DNA, Complementary; D005293:Ferritins; D001052:Apoferritins